This study aims at exploring the potential of ATR-far-ultraviolet (FUV) spectroscopy in investigating electronic structure and transitions of various kinds of biological molecules. For this purpose, ATR-FUV spectra were measured for several kinds of proteins with the different secondary structures, several kinds of carbohydrates, nucleic acids, and lipids. Band assignments have been made for all kinds of biological molecules investigated based on our previous ATR-FUV studies on n-alkanes, alcohols, esters, and amides. For example, the proteins show a characteristic band near 200 nm due to π-π* transition of amide groups. The position of this band varies a little with the secondary structure of proteins but its intensity changes significantly depending on the secondary structure and solutions. All the carbohydrates studied yielded a band near 170 nm due to n-Rydberg transition of ether. In addition, acetylcarbohydrates give an additional band near 190 nm originating from π-π* transition of amide at 2’ carbon. The present study has demonstrated that ATR-FUV spectroscopy is a new powerful technique in exploring electronic structure and transitions of biological molecules, in general. It is also possible to use ATR-FUV spectroscopy for quantitative and qualitative analysis of biological molecules. Moreover, it is of note that information regarding electronic transitions collected by ATR-FUV spectroscopy is useful for UV resonance Raman (UVRR) spectroscopy studies of biological molecules. A combined ATR-FUV spectroscopy and UVRR spectroscopy method may provide a novel analytical tool for molecular and electronic structure of biological molecules.
The combination of confocal micro-Raman spectroscopy and multivariate analysis is carried out for analysis of
maturation of neurons. This study suggests that Raman data reflects the stages of neural maturation which relates with
the expression of new neural function such as spontaneous activity. Neurons obtained from a hippocampus of rat
embryos are cultured in a dish with quartz bottom. According to the previous electrophysiological study, matured neural
cell network showed regulated pulsation with interval of several seconds without any stimulation. It suggested that
alterations in the molecular composition took place in the cell. The Raman measurements are carried out to observe this
alteration along with the maturing process of neurons. The spectra of live neural cells measured after 2, 8, 15, 30, 45, 60,
75, 90, 105 and 120 days of culturing are analyzed by principal component analysis (PCA). The result shows several
groups suggesting the maturation scheme which is observed by the electrophysiological studies. It demonstrates that the
maturation process of neural cells can be monitored by Raman spectroscopy.
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